Measuring Change in Secondary Structure and Oligomeric State for a mAb in Stress and In-Process Testing with MMS and SEC 

21 Nov, 2022 | Newsletters
Measuring Change in Secondary Structure and Oligomeric State for a mAb in Stress and In-Process Testing with MMS and SEC 
Microfluidic Modulation Spectroscopy (MMS) is a new type of Mid-Infrared (MIR) spectroscopy with better sensitivity than traditional MIR and the ability to automate the measurement of samples over a large concentration range. In this talk, we examine the use of MMS to determine the secondary structure of stressed and in-process mAb samples and link these changes to aggregation of the target mAb as seen by size-exclusion chromatography (SEC).

Speaker: Daniel Myatt, PhD – Senior Scientist, Centre for Process Innovation (UK)  
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A Structural Characterisation of a Monoclonal Antibody Therapeutic (Trastuzumab) as Formulated and Under Multiple Conjugation Paradigms
In this study, the commercial mAb Trastuzumab was characterized by MMS directly in its formulation buffer to understand the stand-alone higher-order structure for this important mAb therapeutic. In addition, Trastuzumab was characterized at a variety of different drug-antibody ratios utilizing both non-specific labeling and click-chemistries for the drug conjugation to understand their effects on the mAb structure. Finally, the MMS results were compared to other biophysical techniques such as DSC.
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NEW Behind the App Note Video
Detection of Pressure-Induced Protein Aggregation Using Microfluidic Modulation Spectroscopy (MMS)
 
The ability to detect protein aggregation is important at all stages of drug development.  Early detection of protein aggregation is most desirable to inform development decisions, as it is a recognized signal of instability and can lead to the loss of protein function. Pressure, a stressor used for generating aggregates by impacting noncovalent interactions without the need to change temperature or solvents, was employed to create aggregated human gamma-globulin for this spiked study.‍

​​​​​Dr. Valerie Ivancic Collins, Applications Manager at RedShiftBio hosts this video to highlight how ultra-sensitive MMS was used to obtain aggregation measurements that were previously undetectable by other methods and can greatly influence the direction of biopharma product development:  Significant changes present in the spectral regions of 1624 and 1640cm-1 in the Second Derivative plot that can be seen only subtly in the Absolute Absorbance spectral plot The use of Weighted Spectral Difference (WSD) as a metric to monitor change in sample secondary structure  The strong linear correlation between increasing amounts of percent aggregation and increasing amounts of measured antiparallel beta-sheet content  
 
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